cnwokemodoih's blog

Another study done on plants gives a different dimension to GDSL esterase activity . In this study, a forward genetic screen reveals a brittle leaf sheath 1 (bs1) mutant that is characterized by brittle leaf sheaths. Further genomic investigation, reveals that the BS1 gene codes for a GDSL esterase that removes acetyl moieties from xylan backbones. Because xylan is a major component of secondary cell walls, secondary cell wall architecture is compromised in bs1 mutants. This suggests that this GDSL esterase allows proper secondary wall patterning to occur. BS1 is ubiquitously expressed in plants but more in vascular tissues. Subcellular examination revealed localization in the Golgi apparatus. This is interesting because prior to this discovery, no GDSL esterase/lipase had been discovered to have polysaccharide esterase activity.

The idea of GDSL-lipases/esterases as modulators of immunity is relevant not just in Oryza sativa but also Arabidopsis and hot pepper (Gao et al., 2017). As such, it won’t be out of place to predict that our protein is involved in the modulation of immunity. The plant and subcellular localization patterns are also interesting because they suggest that the same might be the case for our protein. However, we can not draw conclusively on where our protein is expressed; it might be different.

A novel GDSL-type esterase, SFAR4 was found in Arabidopsis thaliana. In this study, knockout mutants and overexpression lines were established. The absence and overexpression of the SFAR4 gene had no effect on germination rate when compared with the wild-types. However, in the presence of glucose, the lines with overexpressed SFAR4 had significantly higher germination rates than wild-type and knockout mutants. In contrast, the presence of mannitol did not affect germination rates. This finding suggests that perhaps SFAR4, a GDSL-type esterase plays a role in glucose susceptibility and glucose metabolic pathway regulation in germinating seeds. In this same study, the expression of fatty acid metabolic genes are shown to be differential in SFAR4 overexpression lines and knockout mutants. SFAR4 overexpression lines show increases levels of metabolism pathway components while knockout mutant lines are accompanied by lower levels of fatty acid metabolism gene expression. This indicates that SFAR4 plays a role in fatty acid metabolism.

Chemical transformation of E. Coli cells is the process by which bacterial cell asre made to take up desired plasmids. Chemical transformation is performed after TOPO cloning. Some preparatory steps include equilibrating the water bath to 42⁰C, warming selective plates at 72⁰C for 30 minutes and thawing vials of One Shot Top 10 E.Coli cells on ice. To start off, 2 microliters of TOPO cloning reaction products are added into vials, mixed gently and then incubated on ice for 5-30 minutes. The cells are heat shocked for 30 second at 42⁰C without shaking before being placed on ice for 2 minutes. 250 microliters of S.O.C. medium is added, the tube is capped tightly and then shook horizontally at 200RPM and 37⁰C for 1 hour. 

The aim of the project is to gain more insight into the function of the alpha-4 GABAA receptor by the deep examination of subunit expression in the brain and spinal cord of zebrafish. The in-situ hybridization procedure done previously (Monesson-Olson et al., 2018), allowed the visualization of six subunit expression patterns, including that of alpha-4. This gave a rough picture of subunit function. Since the mutation screen revealed interesting alpha-4 subunit activity, further analysis is required. Conducting this fluorescent in-situ hybridization procedure will allow me to identify not just where the alpha-4 subunit is expressed but also what structures it interacts with. Knowing what the alpha-4 subunit interacts with will aid in predicting its function. By extension, this will provide a framework for developing treatments that can suppress the effects of related neurological disorders like epilepsy.

Topocloning is a faster and more efficient version of the traditional cloning method. Cloning is the process by which a DNA sequence of interest is integrated into a vector plasmid. This is done traditionally by cutting the plasmid with a restriction enzyme and then ligating either end of the sequence to the open ends of the plasmid. The ligation step is performed by an enzyme called ligase. The use of two different enzymes in two different steps makes the traditional cloning method ineffective. Many errors occur using this method. The advent of topocloning has circumvented the use of two enzymes and reduced errors, by the use of a single enzyme called topoisomerase 1. The vector comes open with two toposiomerase 1 enzymes on either end of the open vector. Once the sequence of interest comes into the gap, the topoisomerase 1 enzymes catalyze the ligation step, closing up the vector and integrating the appropriate sequence into the vector.

Topocloning is a faster and more efficient version of  traditional cloning. Cloning is the process by which a DNA sequence of interest is integrated into a vector plasmid. This is done traditionally by cutting the plasmid with a restriction enzyme and then ligating either end of the sequence to the open ends of the plasmid. The ligation step is typically performed by an enzyme called ligase. The use of two different enzymes in two different steps makes the traditional cloning method ineffective. Many errors occur using this method. The advent of topocloning has circumvented the use of two enzymes, by the use of a single enzyme called topoisomerase 1. The vector comes open with two toposiomerase 1 enzymes on either end of the open vector. Once the sequence of interest comes into the gap, the topoisomerase 1 enzymes catalyze the ligation step, closing up the vector and integrating the appropriate sequence into the vector.

Lawns create the feeling of a natural environment in urban areas, offsetting the health liabilities that typically befall city residents (Irvine et al., 2003), essentially contributing to the overall well-being of the people. Urban green spaces also serve as grounds for recreation and the development of social connection. As for the presence of green spaces on campuses, studies have revealed that there is indeed a correlation between presence and use of green spaces, and perceived student quality of life (McFarland, 2008). This speaks to the fact that lawns or UGSs have vital psychological benefits to humans. Lawns also possess some ecological significance as they contribute to the conservation of biodiversity in urban settings.    

Biodiversity is an important feature in any ecosystem, and lawns are no exception. The data obtained from this study will offer some insight into the effects of common maintenance practices on lawn biodiversity in Amherst, Massachusetts, while also setting the stage for more research that could be done on the topic. The data will be a useful aid in future decisions both about how lawns should be maintained, and about how these practices can be used such that floral diversity is maintained. It can also offer a framework for efforts to slow the decline of biodiversity in green spaces, for example by highlighting which areas have the most biodiversity and how that correlates with how these areas are maintained. This can allow for individualized tailoring of maintenance practices for each lawn or green space based on the plant species that inhabit it, and also provide information about which areas could benefit from the introduction of new species to increase the floral diversity.

Beyond the ecological aspect of maintaining green spaces, there is also the psychological value that lawns have.

Biodiversity is the number of various species of organisms in a given place. Biodiversity ensures that every organism within an ecosystem has a niche in which it can thrive. Generally, the more biodiversity an area has, the more sustainable it is and the more it is able to recover from disaster. With more variation and numbers, an illness for example could infect one species but it would not severely hurt the ecosystem it is in. Lawns have become pervasive with their growing significance in human experiences. Biodiversity is also a feature that accompanies the presence of lawns because various species of flora and fauna exist in these miniature habitats. Plant diversity on lawns is impacted significantly by human disturbances like mowing and use of herbicides which are characteristic of common maintenance practices. These effects have been studied extensively in urban areas but not so much in small towns like Amherst, Massachusetts. In this study, we will investigate the role of these human disturbances here at the University of Massachusetts Amherst and around. We will start by obtaining relevant data about maintenance practices and carrying out sampling experiments in 6 selected lawns. Analysis and comparison of the obtained data will reveal if, in fact, lawn maintenance practices negatively affect plant species diversity. This will provide a framework and resources for future decision-making with regards to of biodiversity conservation.

Biodiversity is the number of various species of organisms in a given place. Biodiversity ensures that every organism within an ecosystem has a niche in which it can thrive. Generally, the more biodiversity an area has, the more sustainable it is and the more it is able to recover from disaster. With more variation and numbers, an illness for example could infect one species but it would not severely hurt the ecosystem it is in. Lawns have become pervasive with their growing significance in human experiences. Biodiversity is also a feature that accompanies the presence of lawns because various species of flora and fauna exist in these miniature habitats. Plant diversity on lawns is impacted significantly by human disturbances like mowing and use of herbicides which are characteristic of common maintenance practices. These effects have been studied extensively in urban areas but not so much in small towns like Amherst, Massachusetts. In this study, we will investigate the role of these human disturbances here at the University of Massachusetts Amherst and around. We will start by obtaining relevant data about maintenance practices and carrying out sampling experiments in 6 selected lawns. Analysis and comparison of the obtained data will reveal if, in fact, lawn maintenance practices negatively affect plant species diversity. This will provide a framework and resources for future decision-making in terms of biodiversity conservation.